• 规格：20μl质粒
• 启动子：T7/lac
• 复制子：ColE1 ori，F1 ori
• 终止子：T7 terminator
• 质粒分类：大肠杆菌载体；PET系列表达质粒
• 质粒大小：6106bp
• 质粒标签：N-6×His , N-DsbA, N-S, N-EK, N-Thrombin,C-6×His
• 原核抗性：卡那霉素Kan
• 克隆菌株：DH5α
• 表达宿主：BL21(DE3)
• 诱导方式：IPTG或乳糖及其类似物
• 5测序引物：T7:TAATACGACTCACTATAGGG
• 3测序引物：T7-ter:TGCTAGTTATTGCTCAGCGG
• 备注：表达DsbA融合蛋白
• 质粒简介：       The pET-39b(+) vector is designed for expression of DsbA fusion proteins. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the T7 terminator primer .
  
• 质粒图谱1： png/3-16101216255A94.png

质粒序列： 点击查看序列

LOCUS       Exported                6106 bp ds-DNA     circular SYN 10-8-2015
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6106)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2015-8-10  from MLCC
            http://www.fenghbio.cn
FEATURES             Location/Qualifiers
     source          1..6106
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1039..1061
                     /organism="Enterobacteria phage T7"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10760"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(150..173)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /note="8xHis"
                     /translation="HHHHHHHH"
     CDS             complement(252..266)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /note="enterokinase site"
                     /translation="DDDDK"
     CDS             complement(282..326)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(342..359)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(369..386)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(408..1031)
                     /codon_start=1
                     /gene="dsbA"
                     /product="bacterial periplasmic oxidoreductase"
                     /note="DsbA"
                     /translation="MKKIWLALAGLVLAFSASAAQYEDGKQYTTLEKPVAGAPQVLEFF
                     SFFCPHCYQFEEVLHISDNVKKKLPEGVKMTKYHVNFMGGDLGKDLTQAWAVAMALGVE
                     DKVTVPLFEGVQKTQTIRSASDIRDVFINAGIKGEEYDAAWNSFVVKSLVAQQEKAAAD
                     VQLRGVPAMFVNGKYQLNPQGMDTSNMDVFVQQYADTVKYLSEKK"
     RBS             1039..1061
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1076..1100
                     /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(1101..1119)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1432..1509
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             1510..2592
                     /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             3401..3592
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3694..3836
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4022..4610)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             4732..5547
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(5640..6095)
                     /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 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
      181 gcggccgcaa gcttgtcgac ggagctcgaa ttcggatccg atatcgccat ggttgaggag
      241 aagcccgggc tcttgtcgtc gtcatcggta cccagatctg ggctgtccat gtgctggcgt
      301 tcgaatttag cagcagcggt ttctttcata ccaattgcag tactaccgcg tggcaccaga
      361 cccgcggagt gatggtgatg gtgatgacca gaaccactag ttgatccttt tttctcgctt
      421 aagtatttca ctgtatcagc atactgctga acaaaaacat ccatattgct ggtatccata
      481 ccctgcggat tcagctgata tttaccgtta acaaacatcg ccggaacgcc acgcaattgc
      541 acgtcagctg cagctttttc ctgctgagcg accagagatt tcaccacgaa gctgttccac
      601 gccgcgtcgt actcttcacc tttaatacct gcgttgataa atacatcgcg gatatcagaa
      661 gcagaacgaa tggtctgggt tttctgtacg ccttcaaaca gcggaacagt cactttgtct
      721 tccacgccca gcgccatcgc cacagcccat gcctgagtca gatctttgcc caggtcacca
      781 cccatgaagt tgacgtggta tttagtcatc ttcacgcctt ccggcagttt tttcttcaca
      841 ttatcagaaa tatgcagaac ttcttcaaac tgatagcagt gcgggcagaa gaaagagaaa
      901 aactccagca cttgcggcgc gccagctacc ggtttttcca gggtagtgta ctgtttacca
      961 tcttcatact gcgccgccga tgcgctaaac gctaaaacta aaccagccag cgccagccaa
     1021 atctttttca tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat
     1081 tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatcga
     1141 tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     1201 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     1261 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
     1321 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac
     1381 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc
     1441 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     1501 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     1561 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     1621 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     1681 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     1741 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     1801 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     1861 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     1921 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     1981 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     2041 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     2101 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     2161 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     2221 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     2281 accgagtccg ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa
     2341 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     2401 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     2461 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     2521 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     2581 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc
     2641 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     2701 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     2761 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
     2821 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
     2881 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccccacgggt
     2941 gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
     3001 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
     3061 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
     3121 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
     3181 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
     3241 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
     3301 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
     3361 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
     3421 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     3481 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     3541 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3601 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3661 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     3721 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     3781 catatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     3841 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3901 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3961 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4021 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4081 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4141 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4201 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4261 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4321 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4381 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4441 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4501 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4561 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4621 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4681 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     4741 attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta
     4801 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
     4861 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     4921 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     4981 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg
     5041 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     5101 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     5161 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     5221 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     5281 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     5341 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     5401 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     5461 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     5521 catttgatgc tcgatgagtt tttctaagaa ttaattcatg agcggataca tatttgaatg
     5581 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5641 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5701 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5761 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5821 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5881 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5941 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     6001 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     6061 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//