| 出品人: | Fodor,B.D.(2004) |
|---|---|
| 载体名称: | pMHE7 |
| 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: | 5920 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: | T7 tag（N-ter）；FLAG-tag、Strep-tag II (C-ter) |
| 载体抗性: | 链霉素 |
| 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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载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS AY303669 5920 bp DNA circular SYN 10-FEB-2004
DEFINITION Broad host range expression vector pMHE7, complete sequence.
ACCESSION AY303669
VERSION AY303669.1 GI:34766420
REFERENCE 1 (bases 1 to 5920)
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.
TITLE Direct Submission
JOURNAL Submitted (21-MAY-2003) Biophysics, BRC Hungarian Academy of
Sciences, Temesvari, Szeged, Csongrad 6701, Hungary
FEATURES Location/Qualifiers
source 1..5920
/organism="Broad host range expression vector pMHE7"
/mol_type="other DNA"
/db_xref="taxon:244944"
primer_bind 195..213
/note="T7 promoter primer"
regulatory 195..211
/regulatory_class="promoter"
/note="T7 promoter"
misc_feature 212..237
/note="lac operator"
regulatory 268..274
/regulatory_class="ribosome_binding_site"
misc_feature 282..284
/note="start codon of fusion protein"
misc_feature 288..311
/note="FLAG-tag"
misc_feature 297..311
/note="enterokinase cleaveage site"
misc_feature 324..347
/note="Strep-tag II"
misc_feature 348..359
/note="factor-Xa cleaveage site"
primer_bind complement(458..475)
/note="T7 terminator primer"
regulatory 471..518
/regulatory_class="terminator"
/note="T7 terminator"
rep_origin 1150..1812
/note="pBBR1 oriV"
misc_feature 2806..3799
/note="RP4 mob, oriT"
regulatory complement(3889..4004)
/regulatory_class="terminator"
/note="T4 terminator"
gene complement(4264..5055)
/gene="aadA"
CDS complement(4264..5055)
/gene="aadA"
/note="streptomycin resistance"
/codon_start=1
/transl_table=11
/product="aminoglycoside-3:adenyltransferase fusion"
/protein_id="AAQ82542.1"
/db_xref="GI:34766421"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKP
HSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYP
AKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQ
DLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPA
QYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
regulatory 5791..5906
/regulatory_class="terminator"
/note="T4 terminator"
ORIGIN
1 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
61 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
121 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
181 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
241 tctagaaata attttgttta actttaagaa ggagatatac catggacgac tacaaggacg
301 acgacgacaa gctcgagggc aactggagcc atccgcagtt cgagaagatc gagggaaggc
361 ctgtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga tccggctgct
421 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
481 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
541 ggatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat
601 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
661 gactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa
721 acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgaaag
781 caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg
841 ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg
901 aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga tcatttattc
961 tgcctcccag agcctgataa aaacggtgaa tccgttagcg aggtgccgcc ggcttccatt
1021 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
1081 agggcggcga ggcggctaca gccgatagtc tggaacagcg cacttacggg ttgctgcgca
1141 acccaagtgc taccggcgcg gcagcgtgac ccgtgtcggc ggctccaacg gctcgccatc
1201 gtccagaaaa cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc gttcccattc
1261 ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg gctggctggg
1321 cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg
1381 gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc
1441 ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca cgcggtcatt
1501 gaccacgtag gccgacacgg tgccggggcc gttgagcttc acgacggaga tccagcgctc
1561 ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga tgagcttgga
1621 aagtgtcttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca cgaggtgatg
1681 cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct catgcgcttt
1741 gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga tttctctgga
1801 ctgcgtggcc atgcttatct ccatgcggta gggtgccgca cggttgcggc accatgcgca
1861 atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca
1921 attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc aatgagctgt
1981 tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg ccctatatct
2041 agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct tcggcgcggc
2101 tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc ggccttcggc
2161 cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg aggctcgggg
2221 ggcaggcggg cgggcttcgc cttcgactgc ccccactcgc ataggcttgg gtcgttccag
2281 gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc cttccacttg
2341 gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc ccagagaaaa
2401 ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg ggtatgtggt
2461 cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag gcgcagcctg
2521 tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca gccggtggcc
2581 gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga ccgcgcaggg
2641 cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc acgactttgc
2701 gaagcaaagt ctagtgagta tactcaagca ttgagtggcc cgccggaggc accgccttgc
2761 gctgcccccg tcgagccggt tggacaccaa aagggagggg caggcatggc ggcatacgcg
2821 atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg cggccagtct caagcacgcc
2881 taccgcgagc gcgagacgcc caacgctgac gccagcagga cgccagagaa cgagcactgg
2941 gcggccagca gcaccgatga agcgatgggc cgactgcgcg agttgctgcc agagaagcgg
3001 cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg ccagcccgga atggtggaag
3061 tcggccagcc aagaacagca ggcggcgttc ttcgagaagg cgcacaagtg gctggcggac
3121 aagtacgggg cggatcgcat cgtgacggcc agcatccacc gtgacgaaac cagcccgcac
3181 atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc tgtcggccaa ggagttcatc
3241 ggcaacaaag cgcagatgac ccgcgaccag accacgtttg cggccgctgt ggccgatcta
3301 gggctgcaac ggggcatcga gggcagcaag gcacgtcaca cgcgcattca ggcgttctac
3361 gaggccctgg agcggccacc agtgggccac gtcaccatca gcccgcaagc ggtcgagcca
3421 cgcgcctatg caccgcaggg attggccgaa aagctgggaa tctcaaagcg cgttgagacg
3481 ccggaagccg tggccgaccg gctgacaaaa gcggttcggc aggggtatga gcctgcccta
3541 caggccgccg caggagcgcg tgagatgcgc aagaaggccg atcaagccca agagacggcc
3601 cgagaccttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt gaatcgggat
3661 atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagctgct gacggaacag
3721 cgggaagtcc agcgccagaa acaggcccag cgccagcagg aacgcgggcg cgcacatttc
3781 cccgaaaagt gccacctggc ggcgttgtga caatttaccg aacaactccg cggccgggaa
3841 gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatag ctttatgctt
3901 gtaaaccgtt ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg gtccccaatt
3961 aattagtaat ataatctatt aaaggtcatt caaaaggtca tccaccggat cagcttagta
4021 aagccctcgc tagattttaa tgcggatgtt gcgattactt cgccaactat tgcgataaca
4081 agaaaaagcc agcctttcat gatatatctc ccaatttgtg tagggcttat tatgcacgct
4141 taaaaataat aaaagcagac ttgacctgat agtttggctg tgagcaatta tgtgcttagt
4201 gcatctaacg cttgagttaa gccgcgccgc gaagcggcgt cggcttgaac gaattgttag
4261 acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa
4321 ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct gtctagcttc
4381 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
4441 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
4501 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
4561 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
4621 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
4681 ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc
4741 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
4801 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
4861 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
4921 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
4981 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
5041 caccgcttcc ctcatgatgt ttaactttgt tttagggcga ctgccctgct gcgtaacatc
5101 gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc
5161 ccgaggcata gactgtaccc caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
5221 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
5281 cttgcattac agcttacgaa ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg
5341 tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg catttctgtc
5401 ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat tggcggcctt
5461 gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg agatcggaag
5521 acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg ttcgcatcct
5581 cggttttctg gaaggcgagc atcgtttgtt cgcccagctt ctgtatggaa cgggcatgcg
5641 gatcagtgag ggtttgcaac tgcgggtcaa ggatctggat ttcgatcacg gcacgatcat
5701 cgtgcgggag ggcaagggct ccaaggatcg ggccttgatg ttacccgaga gcttggcacc
5761 cagcctgcgc gagcagggga attgatccgg tggatgacct tttgaatgac ctttaataga
5821 ttatattact aattaattgg ggaccctaga ggtccccttt tttattttaa aaattttttc
5881 acaaaacggt ttacaagcat aaagctcaag gagatggcgc
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