pTrcHis2B

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  • Model: PVT0811
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pTrcHis2B

PVT0811      2ug
 

 

pTrcHis2B Information

Promoter: Trc/lac

Replicon: PBR322 ori

Terminator: rrnB TT

Plasmid classification: E. coli vector; pTrc series expression plasmid

Plasmid size: 4404 BP

Plasmid label: C-His, C-Myc

Prokaryotic resistance: ampicillin Amp

Cloning strain: DH5alpha

Culture conditions: 37 C, aerobic, LB

Expression host: Escherichia coli

Culture conditions: 37 C, aerobic, LB

Induction: IPTG or lactose and its analogues

5'Sequencing Primer: pTrcHis-F: GAGTATATTAATGTATCG

3'Sequencing Primer: pTrcHis-R: GATTTAATCTGTATCAGG

Note: pTrc expression plasmid

 

pTrcHis2B Description

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.

 

pTrcHis2B Sequence

LOCUS       Exported                4404 bp ds-DNA     circular SYN 29-JUL-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTrcHis2B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4404)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2016-7-29 from SnapGene Viewer 2.8.1
        
FEATURES             Location/Qualifiers
     source          1..4404
                     /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..542
                     /note="MCS"
     CDS             469..498
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /translation="EQKLISEEDL"
     CDS             514..531
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      757..843
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      935..962
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        982..1073
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             1074..1934
                     /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      2105..2693
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    2879..3019
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     promoter        3205..3282
                     /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             3283..4365
                     /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 gctttctaga acaaaaactc
      481 atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca ttgagtttaa
      541 acggtctcca gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
      601 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
      661 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
      721 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
      781 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
      841 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
      901 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
      961 tttgcgtttc tacaaactct ttttgtttat ttttctaaat acattcaaat atgtatccgc
     1021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     1321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2221 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     2281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     2941 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     3001 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3061 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3121 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa
     3181 ggcgaagcgg catgcattta cgttgacacc atcgaatggt gcaaaacctt tcgcggtatg
     3241 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     3301 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     3361 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     3421 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     3481 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     3541 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     3601 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     3661 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     3721 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     3781 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     3841 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     3901 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     3961 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     4021 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     4081 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtca
     4141 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     4201 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     4261 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4321 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4381 tgtgagttag cgcgaattga tctg
//
 

Caution:
Product is for research use only!

 

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pTrcHis2B,Plasmid pTrcHis2B,pTrcHis2B vector

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