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pMHE5 plasmid
BioVector NTCC質粒載體菌種細胞蛋白抗體基因保藏中心www.biovector.net
| 出品人: | Fodor,B.D.(2004) |
|---|---|
| 載體名稱: | pMHE5 |
| 質粒類型: | 廣宿主載體；細菌表達載體;體外表達載體 |
| 高拷貝/低拷貝: | -- |
| 克隆方法: | 限制性內切酶，多克隆位點 |
| 啟動子: | T7/lac |
| 載體大小: | 5938 bp |
| 5' 測序引物及序列: | T7 promoter 5’TAATACGACTCACTATAGGG 3’ |
| 3' 測序引物及序列: | T7 terminator 5’GCTAGTTATTGCTCAGCGG 3’ |
| 載體標簽: | 6XHiS (N ter)；Flag tag、Strep tag II (C ter) |
| 載體抗性: | 鏈霉素 |
| 克隆菌株: | DH5α 等 |
| 宿主細胞（系）: | 多個菌屬 |
| 備注: | -- |
| 產品目錄號: | -- |
| 組成型/誘導型: | 誘導型 |
載體質粒圖譜和多克隆位點信息
載體簡介
載體序列
LOCUS AY299696 5938 bp DNA circular SYN 10-FEB-2004
DEFINITION Broad host range expression vector pMHE5, complete sequence.
ACCESSION AY299696
VERSION AY299696.1 GI:34334146
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 (16-MAY-2003) Biophysics, BRC Hungarian Academy of
Sciences, Temesvari krt., Szeged, Csongrad 6701, Hungary
FEATURES Location/Qualifiers
source 1..5938
/organism="Broad host range expression vector pMHE5"
/mol_type="genomic DNA"
/db_xref="taxon:244054"
primer_bind 193..211
/note="T7 promoter primer"
regulatory 193..209
/regulatory_class="promoter"
/note="T7 promoter"
misc_feature 210..235
/note="lac operator"
regulatory 266..272
/regulatory_class="ribosome_binding_site"
misc_feature 280..282
/note="fusion protein start codon"
misc_feature 286..303
/note="His-tag"
misc_feature 304..315
/note="Factor Xa cleavage site"
misc_feature 370..393
/note="FLAG-tag"
misc_feature 379..393
/note="enterokinase cleavage site"
misc_feature 397..420
/note="Strep-tag II"
primer_bind complement(474..491)
/note="T7 terminator primer"
regulatory 487..534
/regulatory_class="terminator"
/note="T7 terminator"
rep_origin 1166..1828
/note="pBBR1 oriV"
misc_feature 2822..3815
/note="RP4 mob; oriT"
regulatory complement(3905..4020)
/regulatory_class="terminator"
/note="T4 terminator"
gene complement(4280..5071)
/gene="aadA"
CDS complement(4280..5071)
/gene="aadA"
/note="streptomycin resistance"
/codon_start=1
/transl_table=11
/product="aminoglycoside-3:adenyltransferase fusion"
/protein_id="AAQ64667.1"
/db_xref="GI:34334147"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKP
HSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYP
AKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQ
DLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPA
QYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
regulatory 5807..5922
/regulatory_class="terminator"
/note="T4 terminator"
ORIGIN
1 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
61 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
121 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
181 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
241 tagaaataat tttgtttaac tttaagaagg agatatacca tggggcatca tcatcatcat
301 catatcgagg gaaggcctgt cgacggtatc gataagcttg atatcgaatt cctgcagctc
361 gagggatccg actacaagga cgacgacgac aagaactgga gccatccgca gttcgagaag
421 tgaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
481 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
541 aggaggaact atatccggat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
601 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
661 ttttaaccaa taggccgact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
721 ttattggtgc ccttaaacgc ctggtgctac gcctgaataa gtgataataa gcggatgaat
781 ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt cgttaaatag
841 ccgcttatgt ctattgctgg tttaccggtt tattgactac cggaagcagt gtgaccgtgt
901 gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta ataattgacg
961 atatgatcat ttattctgcc tcccagagcc tgataaaaac ggtgaatccg ttagcgaggt
1021 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg
1081 gaggcagaca aggtataggg cggcgaggcg gctacagccg atagtctgga acagcgcact
1141 tacgggttgc tgcgcaaccc aagtgctacc ggcgcggcag cgtgacccgt gtcggcggct
1201 ccaacggctc gccatcgtcc agaaaacacg gctcatcggg catcggcagg cgctgctgcc
1261 cgcgccgttc ccattcctcc gtttcggtca aggctggcag gtctggttcc atgcccggaa
1321 tgccgggctg gctgggcggc tcctcgccgg ggccggtcgg tagttgctgc tcgcccggat
1381 acagggtcgg gatgcggcgc aggtcgccat gccccaacag cgattcgtcc tggtcgtcgt
1441 gatcaaccac cacggcggca ctgaacaccg acaggcgcaa ctggtcgcgg ggctggcccc
1501 acgccacgcg gtcattgacc acgtaggccg acacggtgcc ggggccgttg agcttcacga
1561 cggagatcca gcgctcggcc accaagtcct tgactgcgta ttggaccgtc cgcaaagaac
1621 gtccgatgag cttggaaagt gtcttctggc tgaccaccac ggcgttctgg tggcccatct
1681 gcgccacgag gtgatgcagc agcattgccg ccgtgggttt cctcgcaata agcccggccc
1741 acgcctcatg cgctttgcgt tccgtttgca cccagtgacc gggcttgttc ttggcttgaa
1801 tgccgatttc tctggactgc gtggccatgc ttatctccat gcggtagggt gccgcacggt
1861 tgcggcacca tgcgcaatca gctgcaactt ttcggcagcg cgacaacaat tatgcgttgc
1921 gtaaaagtgg cagtcaatta cagattttct ttaacctacg caatgagcta ttgcgggggg
1981 tgccgcaatg agctgttgcg tacccccctt ttttaagttg ttgattttta agtctttcgc
2041 atttcgccct atatctagtt ctttggtgcc caaagaaggg cacccctgcg gggttccccc
2101 acgccttcgg cgcggctccc cctccggcaa aaagtggccc ctccggggct tgttgatcga
2161 ctgcgcggcc ttcggccttg cccaaggtgg cgctgccccc ttggaacccc cgcactcgcc
2221 gccgtgaggc tcggggggca ggcgggcggg cttcgccttc gactgccccc actcgcatag
2281 gcttgggtcg ttccaggcgc gtcaaggcca agccgctgcg cggtcgctgc gcgagccttg
2341 acccgccttc cacttggtgt ccaaccggca agcgaagcgc gcaggccgca ggccggaggc
2401 ttttccccag agaaaattaa aaaaattgat ggggcaaggc cgcaggccgc gcagttggag
2461 ccggtgggta tgtggtcgaa ggctgggtag ccggtgggca atccctgtgg tcaagctcgt
2521 gggcaggcgc agcctgtcca tcagcttgtc cagcagggtt gtccacgggc cgagcgaagc
2581 gagccagccg gtggccgctc gcggccatcg tccacatatc cacgggctgg caagggagcg
2641 cagcgaccgc gcagggcgaa gcccggagag caagcccgta gggcgccgca gccgccgtag
2701 gcggtcacga ctttgcgaag caaagtctag tgagtatact caagcattga gtggcccgcc
2761 ggaggcaccg ccttgcgctg cccccgtcga gccggttgga caccaaaagg gaggggcagg
2821 catggcggca tacgcgatca tgcgatgcaa gaagctggcg aaaatgggca acgtggcggc
2881 cagtctcaag cacgcctacc gcgagcgcga gacgcccaac gctgacgcca gcaggacgcc
2941 agagaacgag cactgggcgg ccagcagcac cgatgaagcg atgggccgac tgcgcgagtt
3001 gctgccagag aagcggcgca aggacgctgt gttggcggtc gagtacgtca tgacggccag
3061 cccggaatgg tggaagtcgg ccagccaaga acagcaggcg gcgttcttcg agaaggcgca
3121 caagtggctg gcggacaagt acggggcgga tcgcatcgtg acggccagca tccaccgtga
3181 cgaaaccagc ccgcacatga ccgcgttcgt ggtgccgctg acgcaggacg gcaggctgtc
3241 ggccaaggag ttcatcggca acaaagcgca gatgacccgc gaccagacca cgtttgcggc
3301 cgctgtggcc gatctagggc tgcaacgggg catcgagggc agcaaggcac gtcacacgcg
3361 cattcaggcg ttctacgagg ccctggagcg gccaccagtg ggccacgtca ccatcagccc
3421 gcaagcggtc gagccacgcg cctatgcacc gcagggattg gccgaaaagc tgggaatctc
3481 aaagcgcgtt gagacgccgg aagccgtggc cgaccggctg acaaaagcgg ttcggcaggg
3541 gtatgagcct gccctacagg ccgccgcagg agcgcgtgag atgcgcaaga aggccgatca
3601 agcccaagag acggcccgag accttcggga gcgcctgaag cccgttctgg acgccctggg
3661 gccgttgaat cgggatatgc aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa
3721 gctgctgacg gaacagcggg aagtccagcg ccagaaacag gcccagcgcc agcaggaacg
3781 cgggcgcgca catttccccg aaaagtgcca cctggcggcg ttgtgacaat ttaccgaaca
3841 actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg gtacttgggt
3901 cgatagcttt atgcttgtaa accgttttgt gaaaaaattt ttaaaataaa aaaggggacc
3961 tctagggtcc ccaattaatt agtaatataa tctattaaag gtcattcaaa aggtcatcca
4021 ccggatcagc ttagtaaagc cctcgctaga ttttaatgcg gatgttgcga ttacttcgcc
4081 aactattgcg ataacaagaa aaagccagcc tttcatgata tatctcccaa tttgtgtagg
4141 gcttattatg cacgcttaaa aataataaaa gcagacttga cctgatagtt tggctgtgag
4201 caattatgtg cttagtgcat ctaacgcttg agttaagccg cgccgcgaag cggcgtcggc
4261 ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct ttcacgtagt
4321 ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat
4381 aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc
4441 agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg
4501 acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg
4561 ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct
4621 ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca
4681 gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt
4741 ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa
4801 caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca
4861 aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca
4921 gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta
4981 cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag
5041 tcgatacttc ggcgatcacc gcttccctca tgatgtttaa ctttgtttta gggcgactgc
5101 cctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
5161 cttgctgctt ggatgcccga ggcatagact gtaccccaaa aaaacagtca taacaagcca
5221 tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc
5281 gtgagcgcat acgctacttg cattacagct tacgaaccga acaggcttat gtccactggg
5341 ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag
5401 tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc
5461 aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgccctggc
5521 ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg
5581 aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc cagcttctgt
5641 atggaacggg catgcggatc agtgagggtt tgcaactgcg ggtcaaggat ctggatttcg
5701 atcacggcac gatcatcgtg cgggagggca agggctccaa ggatcgggcc ttgatgttac
5761 ccgagagctt ggcacccagc ctgcgcgagc aggggaattg atccggtgga tgaccttttg
5821 aatgaccttt aatagattat attactaatt aattggggac cctagaggtc ccctttttta
5881 ttttaaaaat tttttcacaa aacggtttac aagcataaag ctcaaggaga tggcgccc
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