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!
Search name
pTrcHis2B,Plasmid pTrcHis2B,pTrcHis2B vector