pTrc99a
PVT0806 2ug
pTrc99a Information
Promoter: Trc/lac
Replicon: pBR322 ori
Terminator: rrnB T2
Plasmid classification: Escherichia coli vector; pTrc series expression plasmid
Plasmid size: 4176bp
Prokaryotic resistance: ampicillin Amp
Cloned strain: Escherichia coli DH5 alpha
Culture conditions: 37 centigrade, aerobic, LB
Expression host: Escherichia coli
Culture conditions: 37 centigrade, aerobic, LB
Inducement: IPTG or lactose and its analogues
5'sequencing primers: pTrcHis-F: GAGGTATATATTAATGTATCG
3'sequencing primers: pTrcHis-R:GATTTAATCTGTATCAGG
Remarks: pTrc expression plasmid
pTrc99a Description
PTrc99a is a pTrc series protein expression plasmid of Escherichia coli.
Publication for PVT0806 pTrc99a
Article: A growth-based screening strategy for engineering the catalytic activity of an oxygen-sensitive formate dehydrogenase
Author: Feilong Li 1 2, Silvan Scheller 1, Michael Lienemann 2
Appl Environ Microbiol . 2024 Aug 28:e0147224. doi: 10.1128/aem.01472-24. Online ahead of print.
PMID: 39194220 DOI: 10.1128/aem.01472-24
pTrc99a Sequence
LOCUS Exported 4176 bp ds-DNA circular SYN 11-9-2015
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4176)
AUTHORS admin
TITLE Direct Submission
JOURNAL Exported 2015-9-11
FEATURES Location/Qualifiers
source 1..4176
/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 270..326
/gene="
"
/note="MCS"
/note="pUC18/19 multiple cloning site"
terminator 529..615
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 707..734
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 754..845
/gene="bla"
/note="AmpR promoter"
CDS 846..1706
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1877..2465
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2651..2791
/note="bom"
/note="basis of mobility region from pBR322"
promoter 2977..3054
/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 3055..4137
/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 cagaccatgg aattcgagct cggtacccgg ggatcctcta
301 gagtcgacct gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc
361 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
421 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
481 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
541 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
601 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
661 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
721 acggatggcc tttttgcgtt tctacaaact ctttttgttt atttttctaa atacattcaa
781 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
841 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
901 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
961 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
1021 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
1081 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
1141 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
1201 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
1261 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
1321 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
1381 cgatgcctac agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
1441 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
1501 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
1561 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
1621 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
1681 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
1741 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
1801 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
1861 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
1921 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
1981 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
2041 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
2101 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
2161 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
2221 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
2281 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
2341 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
2401 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
2461 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
2521 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
2581 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
2641 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
2701 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
2761 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
2821 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
2881 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa
2941 ttcgcgcgcg aaggcgaagc ggcatgcatt tacgttgaca ccatcgaatg gtgcaaaacc
3001 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
3061 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
3121 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
3181 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
3241 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
3301 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
3361 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
3421 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
3481 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
3541 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
3601 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
3661 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
3721 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
3781 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
3841 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
3901 atcccgccgt caaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
3961 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
4021 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
4081 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
4141 caacgcaatt aatgtgagtt agcgcgaatt gatctg
//
Caution:
- This product is FOR RESEARCH USE ONLY!
- The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.Take 2ul plasmid into 100 μ L corresponding competent cell, centrifugal then full coating on plate.
- Shipping temperature is 2-8℃.
Search name
pTrc99a,Plasmid pTrc99a,pTrc99a vector
