Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pTrc-fumB | Antibiotic Resistance | Ampicillin |
| Length | 5800 bp | Type | Cloning vector |
| Source | Kim B, Binkley R, Kim HU, Lee SY. | ||
pTrc-fumB vector Vector Map
Plasmid Resuspension protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pTrc-fumB vector Sequence
LOCUS Exported 5800 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTrc-fumB, complete sequence.
ACCESSION MH488916
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5800)
AUTHORS Kim B, Binkley R, Kim HU, Lee SY.
TITLE Metabolic engineering of Escherichia coli for the enhanced
production of l-tyrosine
JOURNAL Biotechnol. Bioeng. (2018) In press
PUBMED 30019750
REFERENCE 2 (bases 1 to 5800)
AUTHORS Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
TITLE Direct Submission
JOURNAL Submitted (15-JUN-2018) Dept. Chemical and Biomolecular Engineering,
Korea Advanced Institute of Science and Technology, Daehak-ro 291,
Daejeon 34141, South Korea
REFERENCE 3 (bases 1 to 5800)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5800
/organism="Cloning vector pTrc-fumB"
/lab_host="Escherichia coli"
/mol_type="other DNA"
/db_xref="taxon:2301177"
regulatory 193..266
/regulatory_class="promoter"
/note="Trc promoter"
promoter 193..222
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 230..246
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
CDS 267..1913
/codon_start=1
/gene="fumB"
/product="fumarase"
/label=fumB
/note="derived from Escherichia coli BL21"
/protein_id="AXN69976.1"
/translation="MSNKPFIYQAPFPMGKDNTEYYLLTSDYVSVADFDGETILKVEPE
ALTLLAQQAFHDASFMLRPAHQKQVAAILHDPEASENDKYVALQFLRNSEIAAKGVLPT
CQDTGTAIIVGKKGQRVWTGGGDEEALSKGVYNTYIEDNLRYSQNAALDMYKEVNTGTN
LPAQIDLYAVDGDEYKFLCVAKGGGSANKTYLYQETKALLTPGKLKNFLVEKMRTLGTA
ACPPYHIAFVIGGTSAETNLKTVKLASAHYYDELPTEGNEHGQAFRDVQLEQELLEEAQ
KLGLGAQFGGKYFAHDIRVIRLPRHGASCPVGMGVSCSADRNIKAKINREGIWIEKLEH
NPDQYIPQELRQAGEGEAVKVDLNRPMKEILAQLSQYPVSTRLSLTGTIIVGRDIAHAK
LKELIDAGKELPQYIKDHPIYYAGPAKTPAGYPSGSLGPTTAGRMDSYVDLLQSHGGSM
IMLAKGNRSQQVTDACHKHGGFYLGSIGGPAAVLAQQSIKHLECVAYPELGMEAIWKIE
VEDFPAFILVDDKGNDFFQQIVNKQCANCTK"
gene 267..1913
/gene="fumB"
/label=fumB
regulatory 2033..2358
/regulatory_class="terminator"
/note="rrnBT1T2"
terminator 2153..2239
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2331..2358
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 2378..2469
/gene="bla"
/label=AmpR promoter
CDS 2470..3330
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/protein_id="AXN69977.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 2470..3330
/gene="bla"
/label=bla
rep_origin 3485..4104
/label=pBR322
/note="pBR322"
rep_origin 3501..4089
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 4275..4415
/label=bom
/note="basis of mobility region from pBR322"
promoter 4601..4678
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 4670..5761
/codon_start=1
/gene="lacIq"
/product="lac repressor protein"
/label=lacIq
/protein_id="AXN69978.1"
/translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
SSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
SPRALADSLMQLARQVSRLESGQ"
gene 4670..5761
/gene="lacIq"
/label=lacIq
CDS 4679..5761
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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"
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