质粒名称: | pmal_c5x_RNAse_Inhib |
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出品公司: | Addgene |
目录编号: | 153314 |
存储实验室: | Drew Endy, Philippa Marrack |
载体骨架基本信息 |
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载体名称: | pMAL-c5X |
载体出品公司: | NEB |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 细菌表达 |
筛选标记: | -- |
载体抗性: | 氨苄霉素 |
生长温度: | 30 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | RNAse Inhibitor |
物种来源: | M. musculus (mouse) |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | Maltose Binding Protein(主链N端) |
克隆方法: | -- |
5’克隆位点: | -- |
3’克隆位点: | -- |
5’测序引物 | -- |
3’测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pmal_c5x_RNAse_Inhib was a gift from Drew Endy & Philippa Marrack (Addgene plasmid # 153314 ; http://n2t.net/addgene:153314 ; RRID:Addgene_153314) |
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 pmal_c5x_RNAse_Inhib 6994 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of an RNase inhibitor. ACCESSION . VERSION . KEYWORDS pmal_c5x_RNAse_Inhib SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6994) TITLE COVID-19 Diagnostic Toolkit Enzymes REFERENCE 2 (bases 1 to 6994) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6994 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 152..180 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 188..204 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 193..215 /label=M13/pUC Reverse /note="In lacZ gene" CDS 274..1374 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /label=MBP /note="This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol." /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF WYAVRTAVINAASGRQTVDEALKDAQT" primer_bind 1348..1371 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 1423..1434 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" terminator 2826..2912 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3004..3031 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 3059..3149 /gene="bla" /label=AmpR promoter CDS 3150..4010 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3368..3387) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4098..4686 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4587..4606 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4840..4857 /label=L4440 /note="L4440 vector, forward primer" primer_bind 5040..5062 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(5056..5247) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" promoter 5743..5820 /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 5821..6903 /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(5840..5859) /label=LacI-R /note="LacI, reverse primer" ORIGIN 1 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat 61 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata 121 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg 181 tgtggaattg tgagcggata acaatttcac acaggaaaca gccagtccgt ttaggtgttt 241 tcacgagcaa ttgaccaaca aggaccatag attatgaaaa tcgaagaagg taaactggta 301 atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag 361 aaagataccg gaattaaagt caccgttgag catccggata aactggaaga gaaattccca 421 caggttgcgg caactggcga tggccctgac attatcttct gggcacacga ccgctttggt 481 ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag 541 ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc 601 gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc 661 tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg 721 ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg 781 ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg 841 aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc 901 gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc 961 ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg 1021 accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc 1081 gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa 1141 ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag 1201 gaagagttgg tgaaagatcc gcgtattgcc gccactatgg aaaacgccca gaaaggtgaa 1261 atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc 1321 aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg 1381 agctcgaaca acaacaacaa taacaataac aacaacctcg ggattgaggg aaggagtctt 1441 gacatccagt gtgagcagct gagtgatgcc cggtggacag agctccttcc tctgatccag 1501 caatacgaag tggtcaggct ggatgactgt ggcctcactg aagtgcggtg caaagacatc 1561 agctcagcag tccaagccaa ccctgccctg acagagctca gccttcgcac caatgaactg 1621 ggtgatggcg gtgtgggtct ggtgctccag ggcctgcaga atcccacttg taagatccag 1681 aagctgagcc ttcagaactg cggcttgacg gaggctggct gtgggatcct gcctggcatg 1741 ctgcgctctt tgtctaccct gcgtgagcta catctcaatg ataaccctat gggggacgca 1801 ggcctgaagc tgctctgtga aggacttcag gacccccagt gccgtcttga gaagcttcag 1861 ttggaatact gtaacctcac agctaccagc tgcgagcccc tggcttcagt gctcagggtg 1921 aaagctgact ttaaggagct agtattgagc aacaacgacc tccatgaacc tggtgtccgt 1981 attctgtgcc agggcctgaa ggactctgcc tgtcaactgg agtcactcaa gctggagaac 2041 tgtggtatca cagcagccaa ctgcaaggat ctgtgcgatg ttgtggcctc caaagcttcg 2101 ctacaggaac tggacttgag cagcaacaag ctaggtaatg caggcattgc agcactgtgc 2161 ccaggactgc tgcttcccag ctgtaagctc aggactctgt ggctctggga atgtgatatc 2221 actgcagagg gctgcaagga cctgtgccgt gtcctcagag ccaagcagag cctgaaggaa 2281 ctcagccttg cttccaatga gctgaaggat gagggtgccc ggctgctgtg tgagagccta 2341 ttagagcctg gctgtcagct ggagtcactg tggataaaga cctgtagcct cacagctgcc 2401 tcctgtccct acttctgctc cgtgttgacc aaaagccgtt ctctgttgga gctgcaaatg 2461 agcagcaacc cactgggaga cgagggagtc caggagcttt gcaaggccct gagccagcca 2521 gatacagtgc tgcgtgagct ttggttggga gactgtgatg tgacaaatag tggttgcagc 2581 agccttgcca atgtcctgct ggccaaccgc agcctgaggg aactggacct cagtaacaac 2641 tgcatggggg gcccaggtgt cctgcaactg ctggagagcc tcaaacagcc cagctgcacc 2701 cttcagcagc tcgtcctgta tgacatttac tggacaaatg aggtggaaga acagcttcgg 2761 gccctggagg aggaaaggcc ttccctgagg atcatttcct aacctgcagg taattaaata 2821 agcttcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt 2881 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga 2941 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta 3001 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt tcggtccgtt 3061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 3121 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 3181 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 3241 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 3301 gcggtaagat ccttgagagt tttcgccccg aagaacgttt cccaatgatg agcactttta 3361 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc 3421 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 3481 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 3541 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 3601 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 3661 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 3721 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 3781 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 3841 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 3901 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 3961 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 4021 aagtttactc atatatactt tagattgatt tccttaggac tgagcgtcaa ccccgtagaa 4081 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4201 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 4261 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 4321 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 4381 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 4441 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 4501 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 4561 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 4621 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 4681 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 4741 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 4801 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 4861 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 4921 atataaggtg cactgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga 4981 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 5041 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg 5101 gtaaagctca tcagcgtggt cgtgcagcga ttcacagatg tctgcctgtt catccgcgtc 5161 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt 5221 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat 5281 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga 5341 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga 5401 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc 5461 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga 5521 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca 5581 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt 5641 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat 5701 catgcgcacc cgtggccagg acccaacgct gcccgaaatt ccgacaccat cgaatggtgc 5761 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat 5821 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt 5881 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg 5941 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag 6001 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc 6061 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa 6121 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt 6181 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc 6241 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt 6301 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag 6361 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc 6421 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg 6481 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact 6541 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc 6601 gggctgcgcg ttggtgcgga tatttcggta gtgggatacg acgataccga agacagctca 6661 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc 6721 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc 6781 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cacaaaccgc ctctccccgc 6841 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 6901 tgagcgcaac gcaattaatg taagttagct cactcattag gcacaattct catgtttgac 6961 agcttatcat cgactgcacg gtgcaccaat gctt //
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