出品公司: | Riggs P. |
---|---|
载体名称: | pMal-c5E |
质粒类型: | 大肠杆菌载体 |
高拷贝/低拷贝: | 高拷贝 |
克隆方法: | -- |
启动子: | Tac |
载体大小: | 5680 bp |
5' 测序引物及序列: | -- |
3' 测序引物及序列: | -- |
载体标签: | -- |
载体抗性: | 氨苄青霉素 |
筛选标记: | -- |
克隆菌株: | DH5α 等 |
宿主细胞(系): | -- |
备注: | -- |
产品目录号: | -- |
稳定性: | -- |
组成型/诱导型: | -- |
病毒/非病毒: | -- |
买家导航
pMal-c5E载体 pMalc5E质粒基本信息
pMal-c5E质粒图谱 pMalc5E载体图谱和pMal-c5E载体序列 pMalc5E质粒序列多克隆位点信息
pMal-c5E质粒 pMalc5E载体简介
pMal-c5E质粒序列 pMalc5E载体序列
LOCUS pMAL-c5E 5680 bp ds-DNA circular SYN 19-SEP-2014 DEFINITION Bacterial vector for inducible cytoplasmic expression of maltose-binding protein (MBP) fusions with an enterokinase cleavage site. ACCESSION . VERSION . KEYWORDS pMAL-c5E SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5680) AUTHORS Riggs P. TITLE Expression and purification of maltose-binding protein fusions. JOURNAL Curr Protoc Mol Biol 2001;Chapter 16:Unit16.6. PUBMED 18265133 REFERENCE 2 (bases 1 to 5680) AUTHORS New England Biolabs TITLE Direct Submission JOURNAL Exported Saturday, January 11, 2020 from SnapGene Viewer 3.1.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5680 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" promoter 3..80 /gene="lacI (mutant)" /note="lacIq promoter" /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 81..1163 /codon_start=1 /gene="lacI" /product="lac repressor" /note="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" misc_feature 1406..1434 /note="tac promoter" /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 1442..1458 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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)." CDS 1528..2628 /codon_start=1 /gene="malE (mutated)" /product="maltose binding protein from E. coli" /note="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" CDS 2677..2691 /codon_start=1 /product="enterokinase recognition and cleavage site" /note="enterokinase site" /translation="DDDDK" misc_feature 2691..2765 /note="MCS" /note="multiple cloning site" terminator 2766..2852 /gene="Escherichia coli rrnB" /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2944..2971 /note="rrnB T2 terminator" /note="transcription terminator T2 from the E. coli rrnB gene" promoter 2999..3089 /gene="bla" /note="AmpR promoter" CDS 3090..3950 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 4038..4626 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4996..5187) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" ORIGIN 1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga 61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg 121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa 181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac 241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc 301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg 361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc 421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca 481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga 541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc 601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg 661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag 721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga 781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa 841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatttcggta gtgggatacg 901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc 961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga 1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata 1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag 1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg 1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg 1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt 1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga 1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga 1501 gcaattgacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg 1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat 1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt 1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac 1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat 1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt 1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa 1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac 1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag 2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg 2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac 2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg 2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc 2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt 2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2461 ttggtgaaag atccgcgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg 2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2641 aacaacaaca acaataacaa taacaacaac ctcggggatg acgatgacaa ggtaccgcat 2701 atgtccatgg gcggccgcga tatcgtcgac ggatccgaat tccctgcagg taattaaata 2761 agcttcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt 2821 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga 2881 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta 2941 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt tcggtccgtt 3001 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 3061 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 3121 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 3181 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 3241 gcggtaagat ccttgagagt tttcgccccg aagaacgttt cccaatgatg agcactttta 3301 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc 3361 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 3421 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 3481 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 3541 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 3601 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 3661 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 3721 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 3781 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 3841 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 3901 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 3961 aagtttactc atatatactt tagattgatt tccttaggac tgagcgtcaa ccccgtagaa 4021 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4081 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4141 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 4201 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 4261 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 4321 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 4381 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 4441 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 4501 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 4561 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 4621 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 4681 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 4741 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 4801 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 4861 atataaggtg cactgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga 4921 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 4981 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg 5041 gtaaagctca tcagcgtggt cgtgcagcga ttcacagatg tctgcctgtt catccgcgtc 5101 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt 5161 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat 5221 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga 5281 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga 5341 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc 5401 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga 5461 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca 5521 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt 5581 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat 5641 catgcgcacc cgtggccagg acccaacgct gcccgaaatt //