出品人: | Fodor,B.D.(2004) |
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载体名称: | pMHE5Tc |
质粒类型: | 广宿主载体;细菌表达载体;体外表达载体 |
高拷贝/低拷贝: | -- |
克隆方法: | 限制性内切酶,多克隆位点 |
启动子: | T7/lac |
载体大小: | 6032 bp |
5' 测序引物及序列: | T7 promoter 5’TAATACGACTCACTATAGGG 3’ |
3' 测序引物及序列: | T7 terminator 5’GCTAGTTATTGCTCAGCGG 3’ |
载体标签: | 6XHis tag(N-ter);FLAG-tag、Strep-tag II (C-ter) |
载体抗性: | 四环素 |
克隆菌株: | DH5α 等 |
宿主细胞(系): | 多个菌属 |
备注: | -- |
产品目录号: | -- |
组成型/诱导型: | 诱导型 |
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载体质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS AY299697 6032 bp DNA circular SYN 10-FEB-2004 DEFINITION Broad host range expression vector pMHE5Tc, complete sequence. ACCESSION AY299697 VERSION AY299697.1 GI:34334148 TITLE Modular broad-host-range expression vectors for single-protein and protein complex purification JOURNAL Appl. Environ. Microbiol. 70 (2), 712-721 (2004) PUBMED 14766546 AUTHORS Fodor,B.D. JOURNAL Submitted (16-MAY-2003) Biophysics, BRC Hungarian Academy of Sciences, Temesvari krt., Szeged, Csongrad 6701, Hungary FEATURES Location/Qualifiers source 1..6032 /organism="Broad host range expression vector pMHE5Tc" /mol_type="genomic DNA" /db_xref="taxon:244055" primer_bind 187..205 /note="T7 promoter primer" regulatory 187..203 /regulatory_class="promoter" /note="T7 promoter" misc_feature 204..229 /note="lac operator" regulatory 260..266 /regulatory_class="ribosome_binding_site" misc_feature 274..276 /note="fusion protein start codon" misc_feature 280..297 /note="His-tag" misc_feature 298..309 /note="Factor Xa cleavage site" misc_feature 364..387 /note="FLAG-tag" misc_feature 373..387 /note="enterokinase cleavage site" misc_feature 391..414 /note="Strep-tag II" primer_bind complement(468..485) /note="T7 terminator primer" regulatory 481..528 /regulatory_class="terminator" /note="T7 terminator" rep_origin 1160..1822 /note="pBBR1 oriV" misc_feature 2816..3809 /note="RP4 mob; oriT" regulatory complement(3899..4014) /regulatory_class="terminator" /note="T4 terminator" gene complement(4320..5510) /gene="tet" CDS complement(4320..5510) /gene="tet" /codon_start=1 /transl_table=11 /product="tetracycline resistance protein" /protein_id="AAQ64668.1" /db_xref="GI:34334149" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASH YGVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAG RIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLH APFLAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTV FFIMQLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGE KQAIIAGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQG QLQGSLAALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSR ATST" regulatory 5895..6010 /regulatory_class="terminator" /note="T4 terminator" ORIGIN 1 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 61 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 121 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg 181 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa 241 taattttgtt taactttaag aaggagatat accatggggc atcatcatca tcatcatatc 301 gagggaaggc ctgtcgacgg tatcgataag cttgatatcg aattcctgca gctcgaggga 361 tccgactaca aggacgacga cgacaagaac tggagccatc cgcagttcga gaagtgaaga 421 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata 481 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg 541 aactatatcc ggatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt 601 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa 661 ccaataggcc gactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg 721 gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga 781 aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt 841 atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct 901 caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga 961 tcatttattc tgcctcccag agcctgataa aaacggtgaa tccgttagcg aggtgccgcc 1021 ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca 1081 gacaaggtat agggcggcga ggcggctaca gccgatagtc tggaacagcg cacttacggg 1141 ttgctgcgca acccaagtgc taccggcgcg gcagcgtgac ccgtgtcggc ggctccaacg 1201 gctcgccatc gtccagaaaa cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc 1261 gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg 1321 gctggctggg cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg 1381 tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa 1441 ccaccacggc ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca 1501 cgcggtcatt gaccacgtag gccgacacgg tgccggggcc gttgagcttc acgacggaga 1561 tccagcgctc ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga 1621 tgagcttgga aagtgtcttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca 1681 cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct 1741 catgcgcttt gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga 1801 tttctctgga ctgcgtggcc atgcttatct ccatgcggta gggtgccgca cggttgcggc 1861 accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa 1921 gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc 1981 aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg 2041 ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct 2101 tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc 2161 ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg 2221 aggctcgggg ggcaggcggg cgggcttcgc cttcgactgc ccccactcgc ataggcttgg 2281 gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc 2341 cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc 2401 ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg 2461 ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag 2521 gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca 2581 gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga 2641 ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc 2701 acgactttgc gaagcaaagt ctagtgagta tactcaagca ttgagtggcc cgccggaggc 2761 accgccttgc gctgcccccg tcgagccggt tggacaccaa aagggagggg caggcatggc 2821 ggcatacgcg atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg cggccagtct 2881 caagcacgcc taccgcgagc gcgagacgcc caacgctgac gccagcagga cgccagagaa 2941 cgagcactgg gcggccagca gcaccgatga agcgatgggc cgactgcgcg agttgctgcc 3001 agagaagcgg cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg ccagcccgga 3061 atggtggaag tcggccagcc aagaacagca ggcggcgttc ttcgagaagg cgcacaagtg 3121 gctggcggac aagtacgggg cggatcgcat cgtgacggcc agcatccacc gtgacgaaac 3181 cagcccgcac atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc tgtcggccaa 3241 ggagttcatc ggcaacaaag cgcagatgac ccgcgaccag accacgtttg cggccgctgt 3301 ggccgatcta gggctgcaac ggggcatcga gggcagcaag gcacgtcaca cgcgcattca 3361 ggcgttctac gaggccctgg agcggccacc agtgggccac gtcaccatca gcccgcaagc 3421 ggtcgagcca cgcgcctatg caccgcaggg attggccgaa aagctgggaa tctcaaagcg 3481 cgttgagacg ccggaagccg tggccgaccg gctgacaaaa gcggttcggc aggggtatga 3541 gcctgcccta caggccgccg caggagcgcg tgagatgcgc aagaaggccg atcaagccca 3601 agagacggcc cgagaccttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt 3661 gaatcgggat atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagctgct 3721 gacggaacag cgggaagtcc agcgccagaa acaggcccag cgccagcagg aacgcgggcg 3781 cgcacatttc cccgaaaagt gccacctggc ggcgttgtga caatttaccg aacaactccg 3841 cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatag 3901 ctttatgctt gtaaaccgtt ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg 3961 gtccccaatt aattagtaat ataatctatt aaaggtcatt caaaaggtca tccaccggat 4021 caattcccct gctcgcgcag gctgggtgcc aagctctcgg gtaacatcaa ggcccgatcc 4081 ttggagccct tgccctcccg cacgatgatc gtgccgtgat cgaaatccag atccttgacc 4141 cgcagttgca aaccctcact gatccgccga gatgcgccgc gtgcggctgc tggagatggc 4201 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa 4261 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt 4321 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat 4381 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc 4441 gccgtgacga tcagcggtcc agtgatcgaa gttaggctgg taagagccgc gagcgatcct 4501 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc 4561 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc 4621 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc 4681 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag 4741 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg 4801 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca 4861 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg 4921 aaggctctca agggcatcgg tcgacgctct cccttatgcg actcctgcat taggaagcag 4981 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag 5041 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg 5101 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg 5161 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc 5221 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg 5281 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga 5341 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa 5401 tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca 5461 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc 5521 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagctcgcg cagatcagtt 5581 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataatgtct 5641 aacaattcgt tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac 5701 taagcacata attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag 5761 cgtgcataat aagccctaca caaattggga gatatatcat gaaaggctgg ctttttcttg 5821 ttatcgcaat agttggcgaa gtaatcgcaa catccgcatt aaaatctagc gagggcttta 5881 ctaagctgat ccggtggatg accttttgaa tgacctttaa tagattatat tactaattaa 5941 ttggggaccc tagaggtccc cttttttatt ttaaaaattt tttcacaaaa cggtttacaa 6001 gcataaagct caaggagatg gcgcccaaca gt //
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