纽普生物

pET-28c(+)载体

载体基本信息
载体名称 pET-28c(+) 载体抗性 Kanamycin
载体长度 5367 bp 载体类型 pET & Duet Vectors (Novagen)
载体来源 Novagen (EMD Millipore) 拷贝数 High copy number

pET-28c(+)载体质粒图谱

pET-28c(+)载体质粒图谱

pET-28c(+)载体载体序列

LOCUS       Exported                5367 bp ds-DNA    circular SYN 02-12-2013
DEFINITION  Bacterial vector for expression of N-terminally 6xHis-tagged 
            proteins with a thrombin site. For other reading frames, use 
            pET-28a(+) or pET-28b(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-28c(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5367)
  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..5367
                     /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(205..237)
                     /codon_start=1
                     /product="epitope tag from a T7 major capsid protein"
                     /note="T7 tag"
                     /translation="MASMTGGQQMG"
     CDS             complement(241..258)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(268..285)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(295..297)
                     /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        693..770
                     /gene="lacI"
                     /note="lacI promoter"
                     /note="
                     "
     CDS             771..1853
                     /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             2662..2853
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(3283..3871)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3993..4808
                     /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(4901..5356)
                     /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 tccgacccat ttgctgtcca ccagtcatgc tagccatatg
      241 gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt
      301 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
      361 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg
      421 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
      481 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
      541 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
      601 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
      661 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
      721 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
      781 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
      841 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
      901 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
      961 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     1021 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     1081 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     1141 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     1201 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     1261 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     1321 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     1381 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     1441 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     1501 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     1561 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     1621 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     1681 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     1741 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1801 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     1861 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
     1921 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     1981 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     2041 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     2101 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     2161 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
     2221 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     2281 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     2341 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     2401 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     2461 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     2521 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     2581 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     2641 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     2701 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     2761 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     2821 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2881 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2941 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3001 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat
     3061 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     3121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3721 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgaaca ataaaactgt
     3961 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
     4021 gctctaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     4081 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     4141 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     4201 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     4261 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
     4321 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     4381 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     4441 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     4501 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     4561 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     4621 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     4681 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     4741 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     4801 ttttctaaga attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4861 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt
     4921 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4981 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     5041 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     5101 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     5161 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     5221 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     5281 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     5341 ccgctacagg gcgcgtccca ttcgcca
//

pET-28c(+)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5367
常规描述Bacterial vector for expression of N-terminally 6xHis-tagged proteins with a thrombin site. For other reading frames, use pET-28a(+) or pET-28b(+).
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..5367
来源分子类型other DNA1..5367
来源种属synthetic DNA construct1..5367
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1205..237
编码区备注T7 tag205..237
编码区产物epitope tag from a T7 major capsid protein205..237
编码区翻译序列MASMTGGQQMG205..237
编码区翻译起始位点1241..258
编码区备注thrombin site241..258
编码区产物thrombin recognition and cleavage site241..258
编码区翻译序列LVPRGS241..258
编码区翻译起始位点1268..285
编码区备注6xHis268..285
编码区产物6xHis affinity tag268..285
编码区翻译序列HHHHHH268..285
编码区翻译起始位点1295..297
编码区备注ATG295..297
编码区产物start codon295..297
编码区翻译序列M295..297
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lacI693..770
启动子备注lacI promoter, 693..770
编码区翻译起始位点1771..1853
编码区基因lacI771..1853
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).771..1853
编码区产物lac repressor771..1853
编码区翻译序列MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ771..1853
编码区翻译起始位点12662..2853
编码区基因rop2662..2853
编码区备注rop2662..2853
编码区产物Rop protein2662..2853
编码区翻译序列MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL2662..2853
复制起点方向LEFT3283..3871
复制起点备注ori,high-copy-number ColE1/pMB1/pBR322/pUC origin of replication3283..3871
编码区翻译起始位点13993..4808
编码区基因aph(3')-Ia3993..4808
编码区备注KanR,confers resistance to kanamycin3993..4808
编码区产物aminoglycoside phosphotransferase3993..4808
编码区翻译序列MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF3993..4808
复制起点方向LEFT4901..5356
复制起点备注f1 ori,f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis4901..5356

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