• 商品货号：FH1592

• 规格：10μl质粒
• 启动子：Trc/lac promoter
• 复制子：PBR322 ori
• 终止子：rrnB TT
• 质粒大小：4406bp
• 质粒标签：C-Myc，C-6xHis
• 原核抗性：氨苄青霉素Amp
• 克隆菌株：大肠杆菌DH5α
• 表达宿主：大肠杆菌
• 诱导方式：IPTG或乳糖及其类似物
• 5测序引物：pTrcHis-F: GAGGTATATATTAATGTATCG
• 3测序引物：pTrcHis-F: GAGGTATATATTAATGTATCG
• 备注：pTrc表达质粒
• 质粒简介：          pTrcHis2A质粒是一个大肠杆菌蛋白表达载体，Trc启动子驱动目的基因的表达。The pTrcHis2 plasmids are pBR322-derived expression vectors designed for efficient recombinant protein expression and purification in E. coli. High levels of expression are possible using the trc (trp-lac) promoter (Egon et al., 1983) and the rrnB anti-termination region (Li et al., 1984). The trc promoter contains the –35 region of the trp promoter together with the –10 region of the lac promoter (Brosius et al., 1985; Egon et al., 1983; Mulligan et al., 1985). To regulate expression, the gene encoding Lac repressor (lacIq) is provided in the pTrcHis2 vectors, allowing regulation of the trc promoter regardless of whether the host strain contains a gene encoding the Lac repressor. Isopropyl-Beta-D-thiogalactopyranoside (IPTG) is used to induce expression of your gene. Translation is enhanced by the bacteriophage T7 gene 10 translation enhancer and a minicistron that provides highly efficient translational restart into the open reading frame of the multiple cloning site. DNA inserts are positioned downstream and in frame with the initiation ATG and a C-terminal fusion peptide. The C-terminal peptide encodes the myc epitope and six histidine residues that function as a metal binding site in the expressed protein.
• 质粒图谱1： gif/1-1G0261Q300456.gif

质粒序列： 点击查看序列

LOCUS       Exported                4406 bp ds-DNA     circular SYN 29-七月-2016
DEFINITION  synthetic circular DNA
KEYWORDS    pTrcHis2A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4406)
  AUTHORS   .
  TITLE     Direct Submission
            http://www.fenghbio.cn
FEATURES             Location/Qualifiers
     source          1..4406
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    193..222
                     /note="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     misc_feature    263..332
                     /note="rrnG antiterminator"
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /note="minicistron"
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     misc_feature    411..541
                     /note="MCS"
     CDS             471..500
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /translation="EQKLISEEDL"
     CDS             516..533
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      759..845
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      937..964
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        984..1075
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             1076..1936
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2107..2695
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    2881..3021
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     promoter        3207..3284
                     /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."
     CDS             3285..4367
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatggatcc
      421 gagctcgaga tctgcagctg gtaccatatg ggaattcgaa gcttgggccc gaacaaaaac
      481 tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt
      541 aaacggtctc cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
      601 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
      661 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
      721 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
      781 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
      841 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
      901 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
      961 tttttgcgtt tctacaaact ctttttgttt atttttctaa atacattcaa atatgtatcc
     1021 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1081 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1141 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1201 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1261 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     1321 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1381 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1441 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1501 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1561 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1621 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1681 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1741 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1801 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1861 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1921 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1981 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2101 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2161 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2221 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2281 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2341 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2401 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2461 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2521 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2581 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2641 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2701 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2761 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2821 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2881 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     2941 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     3001 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3061 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3121 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg
     3181 aaggcgaagc ggcatgcatt tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta
     3241 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     3301 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     3361 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     3421 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     3481 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     3541 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     3601 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     3661 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     3721 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     3781 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     3841 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     3901 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     3961 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     4021 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     4081 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     4141 caaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     4201 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     4261 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4321 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4381 aatgtgagtt agcgcgaatt gatctg
//