纽普生物

pET-29a(+)载体

载体基本信息
载体名称 pET-29a(+) 载体抗性 Kanamycin
载体长度 5371 bp 载体类型 pET & Duet Vectors (Novagen)
载体来源 Novagen (EMD Millipore) 拷贝数 High copy number

pET-29a(+)载体质粒图谱

pET-29a(+)载体质粒图谱

pET-29a(+)载体载体序列

LOCUS       Exported                5371 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-29b(+) or 
            pET-29c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-29a(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5371)
  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..5371
                     /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(216..233)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(249..293)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(294..296)
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     RBS             305..310
                     /note="ribosome binding site"
     protein_bind    341..365
                     /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(366..384)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        697..774
                     /gene="lacI"
                     /note="lacI promoter"
                     /note="
                     "
     CDS             775..1857
                     /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             2666..2857
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(3287..3875)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3997..4812
                     /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(4905..5360)
                     /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 tccgatatca gccatggaac cgcgtggcac cagggtaccc
      241 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatatgt
      301 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
      361 attcccctat agtgagtcgt attaatttcg cgggatcgag atcgatctcg atcctctacg
      421 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
      481 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
      541 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
      601 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
      661 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
      721 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
      781 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
      841 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
      901 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
      961 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     1021 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     1081 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     1141 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     1201 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     1261 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     1321 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     1381 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     1441 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     1501 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     1561 cgcgttggtg cggacatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     1621 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     1681 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     1741 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     1801 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     1861 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
     1921 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     1981 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     2041 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     2101 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     2161 cggcgagaag caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct
     2221 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     2281 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     2341 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     2401 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     2461 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     2521 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     2581 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     2641 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     2701 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     2761 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     2821 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2881 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2941 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     3001 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat atgcggtgtg
     3061 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     3121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3721 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg aacaataaaa
     3961 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     4021 tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     4081 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     4141 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     4201 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     4261 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag
     4321 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     4381 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     4441 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     4501 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     4561 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     4621 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     4681 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     4741 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     4801 gagtttttct aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa
     4921 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4981 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5041 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5101 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5161 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5221 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5281 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5341 tgcgccgcta cagggcgcgt cccattcgcc a
//

pET-29a(+)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5371
常规描述Bacterial vector for expression of N-terminally S-tagged proteins with a thrombin site. For other reading frames, use pET-29b(+) 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..5371
来源分子类型other DNA1..5371
来源种属synthetic DNA construct1..5371
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1216..233
编码区备注thrombin site216..233
编码区产物thrombin recognition and cleavage site216..233
编码区翻译序列LVPRGS216..233
编码区翻译起始位点1249..293
编码区备注S-Tag249..293
编码区产物affinity and epitope tag derived from pancreatic ribonuclease A249..293
编码区翻译序列KETAAAKFERQHMDS249..293
编码区翻译起始位点1294..296
编码区备注ATG294..296
编码区产物start codon294..296
编码区翻译序列M294..296
核糖体结合位点备注ribosome binding site305..310
蛋白结合位点嵌合范围lac repressor encoded by lacI341..365
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).341..365
启动子备注T7 promoter,promoter for bacteriophage T7 RNA polymerase366..384
启动子基因lacI697..774
启动子备注lacI promoter, 697..774
编码区翻译起始位点1775..1857
编码区基因lacI775..1857
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).775..1857
编码区产物lac repressor775..1857
编码区翻译序列MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ775..1857
编码区翻译起始位点12666..2857
编码区基因rop2666..2857
编码区备注rop2666..2857
编码区产物Rop protein2666..2857
编码区翻译序列MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL2666..2857
复制起点方向LEFT3287..3875
复制起点备注ori,high-copy-number ColE1/pMB1/pBR322/pUC origin of replication3287..3875
编码区翻译起始位点13997..4812
编码区基因aph(3')-Ia3997..4812
编码区备注KanR,confers resistance to kanamycin3997..4812
编码区产物aminoglycoside phosphotransferase3997..4812
编码区翻译序列MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF3997..4812
复制起点方向LEFT4905..5360
复制起点备注f1 ori,f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis4905..5360

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