Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pTrc-HisTAL | Antibiotic Resistance | Ampicillin |
| Length | 5716 bp | Type | Cloning vector |
| Source | Kim B, Binkley R, Kim HU, Lee SY. | ||
pTrc-HisTAL 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-HisTAL vector Sequence
LOCUS Exported 5716 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTrc-HisTAL, complete sequence.
ACCESSION MH488930
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5716)
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 5716)
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 5716)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5716
/organism="Cloning vector pTrc-HisTAL"
/lab_host="Escherichia coli"
/mol_type="other DNA"
/db_xref="taxon:2301173"
regulatory 193..246
/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)."
5'UTR 247..271
/gene="TAL"
gene 247..271
/gene="TAL"
/label=TAL
CDS 272..1822
/codon_start=1
/gene="TAL"
/product="tyrosine ammonia-lyase"
/label=TAL
/note="N-terminal 6XHis-tagged"
/protein_id="AXN70019.1"
/translation="MHHHHHHTQVVERQADRLSSREYLARVVRSAGWDAGLTSCTDEEI
VRMGASARTIEEYLKSDKPIYGLTQGFGPLVLFDADSELEQGGSLISHLGTGQGAPLAP
EVSRLILWLRIQNMRKGYSAVSPVFWQKLADLWNKGFTPAIPRHGTVSASGDLQPLAHA
ALAFTGVGEAWTRDADGRWSTVPAVDALAALGAEPFDWPVREALAFVNGTGASLAVAVL
NHRSALRLVRACAVLSARLATLLGANPEHYDVGHGVARGQVGQLTAAEWIRQGLPRGMV
RDGSRPLQEPYSLRCAPQVLGAVLDQLDGAGDVLAREVDGCQDNPITYEGELLHGGNFH
AMPVGFASDQIGLAMHMAAYLAERQLGLLVSPVTNGDLPPMLTPRAGRGAGLAGVQISA
TSFVSRIRQLVFPASLTTLPTNGWNQDHVPMALNGANSVFEALELGWLTVGSLAVGVAQ
LAAMTGHAAEGVWAELAGICPPLDADRPLGAEVRAARDLLSAHADQLLVDEADGKDFG"
gene 272..1822
/gene="TAL"
/label=TAL
CDS 275..292
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
regulatory 1949..2274
/regulatory_class="terminator"
/note="rrnBT1T2"
terminator 2069..2155
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2247..2274
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 2294..2385
/gene="bla"
/label=AmpR promoter
CDS 2386..3246
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/protein_id="AXN70020.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 2386..3246
/gene="bla"
/label=bla
rep_origin 3401..4020
/label=pBR322
/note="pBR322"
rep_origin 3417..4005
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 4191..4331
/label=bom
/note="basis of mobility region from pBR322"
promoter 4517..4594
/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 4586..5677
/codon_start=1
/gene="lacIq"
/product="lac repressor protein"
/label=lacIq
/protein_id="AXN70021.1"
/translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
SSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
SPRALADSLMQLARQVSRLESGQ"
gene 4586..5677
/gene="lacIq"
/label=lacIq
CDS 4595..5677
/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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