质粒名称: | pRE112 |
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出品公司: | 爱迪基因 |
目录编号: | 43828 |
存储实验室: | Dieter Schifferli |
载体骨架基本信息 |
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载体名称: | pRE107 (modified from pGP704) |
载体出品公司: | Addgene plasmid 43829 |
载体抗性: | 氯霉素 |
空载体大小: | 5173 bp |
完整质粒大小: | -- |
载体修饰: | 来源于pGP704:SacB1从pUC58-SacB1克隆到EcoRI位点。oriV的BamHI位点经部分消化和Klenow处理转化为ClaI位点。从107年前开始:氨苄西林抗性被氯霉素抗性所取代(来自pACYC184)被钝化的BamHI消化。 |
载体类型: | 细菌表达;sacB1等位基因交换载体 |
筛选标记: | SacB (sucrose sensitivity) |
高拷贝/低拷贝: | 低拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | -- |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | CAT-F |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pRE112 was a gift from Dieter Schifferli (Addgene plasmid # 43828 ; http://n2t.net/addgene:43828 ; RRID:Addgene_43828) |
参考文献部分: | Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression. Edwards RA, Keller LH, Schifferli DM. Gene. 1998 Jan 30;207(2):149-57. 10.1016/S0378-1119(97)00619-7 PubMed 9511756 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pRE112 5760 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRE112 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5760) AUTHORS Edwards RA, Keller LH, Schifferli DM TITLE Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression. JOURNAL Gene. 1998 Jan 30;207(2):149-57. PUBMED 9511756 REFERENCE 2 (bases 1 to 5760) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5760 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 93..482 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" promoter 510..955 /gene="sacR" /label=sacB promoter /note="sacB promoter and control region" CDS 956..2377 /codon_start=1 /gene="Bacillus subtilis sacB" /product="secreted levansucrase that renders bacterial growth sensitive to sucrose" /label=SacB /note="negative selection marker" /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV VKDSILEQGQLTVNK" CDS complement(3216..3875) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 3790..3809 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(3876..3978) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" oriT 4707..4816 /note="incP origin of transfer" CDS 4849..5220 /codon_start=1 /gene="traJ" /product="oriT-recognizing protein" /label=traJ /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP" ORIGIN 1 cgcgtttccg agaaccgcgc gaacgacatg gagcggcacg cgggcgtgga aagcctggtc 61 ggctggatcg gcacgatgcg tccggcgtag aggatctgaa gatccagcag ttcaacctgt 121 tgatagtacg tactaagctc tcatgtttca cgtactaagc tctcatgttt aacgtactaa 181 gctctcatgt ttaacgaact aaaccctcat ggctaacgta ctaagctctc atggctaacg 241 tactaagctc tcatgtttca cgtactaagc tctcatgttt gaacaataaa attaatataa 301 atcagcaact taaatagcct ctaaggtttt aagttttata agaaaaaaaa gaatatataa 361 ggcttttaaa gcttttaagg tttaacggtt gtggacaaca agccagggat gtaacgcact 421 gagaagccct tagagcctct caaagcaatt ttcagtgaca caggaacact taacggctga 481 catgggaatt ctgatccttt ttaacccatc acatatacct gccgttcact attatttagt 541 gaaatgagat attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc 601 aacagaaact ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta 661 ggcccgtagt ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca 721 gttgcaatcc aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt 781 tactgataaa gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca 841 ccccttacat attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag 901 gagggctgga agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa 961 catcaaaaag tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg 1021 aggcgcaact caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg 1081 catttcccat attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa 1141 atatcaagtt cctgagttcg attcgtccac aattaaaaat atctcttctg caaaaggcct 1201 ggacgtttgg gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg 1261 ctaccacatc gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta 1321 catgttctat caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt 1381 ctttaaagac agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga 1441 atggtcaggt tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt 1501 ctccggtaaa cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc 1561 agacagctct ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg 1621 aaaaacgtat caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa 1681 ccatacgctg agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga 1741 agcaaacact ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata 1801 ctatggcaaa agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa 1861 aaaacgcacg gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta 1921 cacactgaaa aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga 1981 acgcgcgaac gtctttaaaa tgaacggcaa atggtatctg ttcactgact cccgcggatc 2041 aaaaatgacg attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa 2101 ttctttaact ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct 2161 tgatcctaac gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa 2221 caatgtcgtg attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac 2281 gtttgcgccc agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag 2341 catccttgaa caaggacaat taacagttaa caaataaaaa cgcaaaagaa aatgccgata 2401 tcctattggc attttctttt atttcttatc aacataaagg tgaatcccat atgaactata 2461 taaaagcagg caaatggcta accgtattcc taaccttttg gtaatgactc caacttattg 2521 atagtgtttt atgttcagat aatgcccgat gactttgtca tgcagctcca ccgattttga 2581 gaacgacagc gacttccgtc ccagccgtgc caggtgctgc ctcagattca ggttatgccg 2641 ctcaattcgc tgcgtatatc gcttgctgat tacgtgcagc tttcccttca ggcgggattc 2701 atacagcggc cagccatccg tcatccatat caccacgtca aagggtgaca gcaggctcat 2761 aagacgcccc agcgtcgcca tagtgcgttc accgaatacg tgcgcaacaa ccgtcttccg 2821 gagactgtca tacgcgtaaa acagccagcg ctggcgcgat ttagccccga catagcccca 2881 ctgttcgtcc atttccgcgc agacgatgac gtcactgccc ggctgtatgc gcgaggttac 2941 cgactgcggc ctgagttttt taagtgacgt aaaatcgtgt tgaggccaac gcccataatg 3001 cgggctgttg cccggcatcc aacgccattc atggccatat caatgatttt ctggtgcgta 3061 ccgggttgag aagcggtgta agtgaactgc atgaattccc gggagagctc gatatcgcat 3121 gcggtacctc tagaagaagc ttgggatcgg gccctatcac ttattcaggc gtagcaacca 3181 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg 3241 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca gacggcatga 3301 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg 3361 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa 3421 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag 3481 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa 3541 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg 3601 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat 3661 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc 3721 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag 3781 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata 3841 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat 3901 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa 3961 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta 4021 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtattagg 4081 gcccgatcct ttttgtccgg tgttgggttg aaggtgaagc cggtcggggc cgcagcgggg 4141 gccggctttt cagccttgcc cccctgcttc ggccgccgtg gctccggcgt cttgggtgcc 4201 ggcgcgggtt ccgcagcctt ggcctgcggt gcgggcacat cggcgggctt ggccttgatg 4261 tgccgcctgg cgtgcgagcg gaacgtctcg taggagaact tgaccttccc cgtttcccgc 4321 atgtgctccc aaatggtgac gagcgcatag ccggacgcta acgccgcctc gacatccgcc 4381 ctcaccgcca ggaacgcaac cgcagcctca tcacgccggc gcttcttggc cgcgcgggat 4441 tcaacccact cggccagctc gtcggtgtag ctctttggca tcgtctctcg cctgtcccct 4501 cagttcagta atttcctgca tttgcctgtt tccagtcggt agatattcca caaaacagca 4561 gggaagcagc gcttttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg 4621 gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt 4681 ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc 4741 gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct 4801 ctgcgaggct ggccggctac cgccggcgta acagatgagg gcaagcggat ggctgatgaa 4861 accaagccaa ccaggaaggg cagcccacct atcaaggtgt actgccttcc agacgaacga 4921 agagcgattg aggaaaaggc ggcggcggcc ggcatgagcc tgtcggccta cctgctggcc 4981 gtcggccagg gctacaaaat cacgggcgtc gtggactatg agcacgtccg cgagctggcc 5041 cgcatcaatg gcgacctggg ccgcctgggc ggcctgctga aactctggct caccgacgac 5101 ccgcgcacgg cgcggttcgg tgatgccacg atcctcgccc tgctggcgaa gatcgaagag 5161 aagcaggacg agcttggcaa ggtcatgatg ggcgtggtcc gcccgagggc agagccatga 5221 cttttttagc cgctaaaacg gccggggggt gcgcgtgatt gccaagcacg tccccatgcg 5281 ctccatcaag aagagcgact tcgcggagct ggtgaagtac atcaccgacg agcaaggcaa 5341 gaccgagcgc ctgggtcacg tgcgcgtcac gaactgcgag gcaaacaccc tgcccgctgt 5401 catggccgag gtgatggcga cccagcacgg caacacccgt tccgaggccg acaagaccta 5461 tcacctgctg gttagcttcc gcgcgggaga gaagcccgac gcggagacgt tgcgcgcgat 5521 tgaggaccgc atctgcgctg ggcttggctt cgccgagcat cagcgcgtca gtgccgtgca 5581 tcacgacacc gacaacctgc acatccatat cgccatcaac aagattcacc cgacccgaaa 5641 caccatccat gagccgtatc gggcctaccg cgccctcgct gacctctgcg cgacgctcga 5701 acgggactac gggcttgagc gtgacaatca cgaaacgcgg cagcgcgttt ccgagaaccg //