纽普生物

pET-29b(+)载体

载体基本信息
载体名称 pET-29b(+) 载体抗性 Kanamycin
载体长度 5370 bp 载体类型 pET & Duet Vectors (Novagen)
载体来源 Novagen (EMD Millipore) 拷贝数 High copy number

pET-29b(+)载体质粒图谱

pET-29b(+)载体质粒图谱

pET-29b(+)载体载体序列

LOCUS       Exported                5370 bp ds-DNA    circular SYN 02-12-2013
DEFINITION  Bacterial vector for expression of N-terminally S-tagged proteins 
            with a thrombin site. For other reading frames, use pET-29a(+) or 
            pET-29c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-29b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5370)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
  JOURNAL   Exported 2017/2/28 from SnapGene 2.3.2
            http://www.snapgene.com
COMMENT     
FEATURES             Location/Qualifiers
     source          1..5370
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     misc_feature    158..203
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(215..232)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(248..292)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(293..295)
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     RBS             304..309
                     /note="ribosome binding site"
     protein_bind    340..364
                     /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)."
     promoter        complement(365..383)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        696..773
                     /gene="lacI"
                     /note="lacI promoter"
                     /note="
                     "
     CDS             774..1856
                     /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"
     CDS             2665..2856
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(3286..3874)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3996..4811
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(4904..5359)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgatatcg ccatggaacc gcgtggcacc agggtaccca
      241 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcatatgta
      301 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
      361 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga tcctctacgc
      421 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
      481 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
      541 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
      601 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
      661 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct
      721 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
      781 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
      841 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
      901 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
      961 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     1021 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     1081 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     1141 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     1201 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     1261 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     1321 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     1381 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     1441 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     1501 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     1561 gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     1621 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     1681 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     1741 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     1801 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     1861 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg
     1921 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     1981 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
     2041 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
     2101 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
     2161 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc
     2221 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     2281 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     2341 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     2401 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     2461 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     2521 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     2581 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     2641 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     2701 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     2761 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
     2821 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2881 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2941 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     3001 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga
     3061 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     3121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3721 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac
     3961 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     4021 cttgctctag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     4081 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
     4141 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     4201 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     4261 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg
     4321 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     4381 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     4441 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     4501 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     4561 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     4621 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     4681 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     4741 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     4801 agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa
     4861 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat
     4921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5341 gcgccgctac agggcgcgtc ccattcgcca
//

pET-29b(+)载体载体特征

一级属性 二级属性 属性值 起始位置
常规长度5370
常规描述Bacterial vector for expression of N-terminally S-tagged proteins with a thrombin site. For other reading frames, use pET-29a(+) or pET-29c(+).
常规是否环形1
常规作者Novagen (EMD Millipore)
常规注释
来源实验宿主Escherichia coli1..5370
来源分子类型other DNA1..5370
来源种属synthetic DNA construct1..5370
终止子备注T7 terminator,transcription terminator for bacteriophage T7 RNA polymerase26..73
编码区翻译起始位点1140..157
编码区备注6xHis140..157
编码区产物6xHis affinity tag140..157
编码区翻译序列HHHHHH140..157
其它特征区备注MCS,multiple cloning site158..203
编码区翻译起始位点1215..232
编码区备注thrombin site215..232
编码区产物thrombin recognition and cleavage site215..232
编码区翻译序列LVPRGS215..232
编码区翻译起始位点1248..292
编码区备注S-Tag248..292
编码区产物affinity and epitope tag derived from pancreatic ribonuclease A248..292
编码区翻译序列KETAAAKFERQHMDS248..292
编码区翻译起始位点1293..295
编码区备注ATG293..295
编码区产物start codon293..295
编码区翻译序列M293..295
核糖体结合位点备注ribosome binding site304..309
蛋白结合位点嵌合范围lac repressor encoded by lacI340..364
蛋白结合位点备注lac operator,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).340..364
启动子备注T7 promoter,promoter for bacteriophage T7 RNA polymerase365..383
启动子基因lacI696..773
启动子备注lacI promoter, 696..773
编码区翻译起始位点1774..1856
编码区基因lacI774..1856
编码区备注lacI,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).774..1856
编码区产物lac repressor774..1856
编码区翻译序列MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ774..1856
编码区翻译起始位点12665..2856
编码区基因rop2665..2856
编码区备注rop2665..2856
编码区产物Rop protein2665..2856
编码区翻译序列MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL2665..2856
复制起点方向LEFT3286..3874
复制起点备注ori,high-copy-number ColE1/pMB1/pBR322/pUC origin of replication3286..3874
编码区翻译起始位点13996..4811
编码区基因aph(3')-Ia3996..4811
编码区备注KanR,confers resistance to kanamycin3996..4811
编码区产物aminoglycoside phosphotransferase3996..4811
编码区翻译序列MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF3996..4811
复制起点方向LEFT4904..5359
复制起点备注f1 ori,f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis4904..5359

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