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名称	规格
pQE-82L-Kan	20μl

启动子：T5

复制子：ColE1 ori

终止子：Lambda t0 terminator；rrnB T1

质粒分类：大肠杆菌载体；pQE系列表达质粒

质粒大小：4542bp

原核抗性：卡那霉素Kan

克隆菌株：DH5α

培养条件：37℃，有氧，LB

表达宿主：M15

培养条件：37℃，有氧，LB

诱导方式：IPTG或乳糖及其类似物

5'测序引物：pQE30-F: AGCGGATAACAATTTCACACAG

3'测序引物：pQE30-R: TTCTGAGGTCATTACTGGATC
 

质粒序列
LOCUS       Exported                4542 bp ds-DNA     circular SYN 05-DEC-2013
DEFINITION  Bacterial lacIq vector with a kanamycin resistance marker for
            expressing N-terminally 6xHis-tagged proteins. For other reading
            frames, use pQE-80L-Kan or pQE-81L-Kan.
ACCESSION   .
VERSION     .
KEYWORDS    pQE-82L-Kan
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4542)
  AUTHORS   Qiagen
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, September 14, 2016 from SnapGene Viewer 3.1.4
COMMENT     Because this vector contains the lacIq gene, the pREP4 plasmid is
            not needed.
FEATURES             Location/Qualifiers
     source          1..4542
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        10..54
                     /note="T5 promoter"
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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)."
     protein_bind    62..78
                     /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)."
     RBS             101..106
                     /note="ribosome binding site"
     CDS             115..117
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     misc_feature    146..193
                     /note="MCS"
                     /note="multiple cloning site"
     misc_feature    193..203
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     terminator      209..303
                     /note="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             347..1006
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      1071..1157
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(1250..2332)
                     /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"
     promoter        complement(2333..2410)
                     /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."
     rep_origin      complement(2928..3516)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3623..4438)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatctcacc atcaccatca ccatgggatc cgcatgcgag ctcggtaccc cgggtcgacc
      181 tgcagccaag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
      241 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      301 aatccaagct agcttggcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
      361 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
      421 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
      481 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
      541 cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg
      601 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
      661 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
      721 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
      781 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
      841 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
      901 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
      961 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
     1021 ttattggtgc ccttaaacgc ctggggtaat gactctctag cttgaggcat caaataaaac
     1081 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1141 tcctgagtag gacaaatccg ccctctagat tacgtgcagt cgatgataag ctgtcaaaca
     1201 tgagaattgt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     1261 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1321 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     1381 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     1441 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     1501 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
     1561 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     1621 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     1681 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     1741 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     1801 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     1861 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     1921 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     1981 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     2041 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     2101 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     2161 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     2221 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     2281 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     2341 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt
     2401 cgatggtgtc ggaatttcgg gcagcgttgg gtcctggcca cgggtgcgca tgatctagag
     2461 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2521 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2581 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     2641 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     2701 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     2761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3121 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3361 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat ggcgttaagg
     3601 gattttggtc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     3661 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     3721 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     3781 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     3841 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     3901 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     3961 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     4021 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     4081 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     4141 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     4201 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     4261 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     4321 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     4381 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     4441 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac catgacatta
     4501 acctataaaa ataggcgtat cacgaggccc tttcgtcttc ac
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